CN103520766A - Mussel mucoprotein liquid product as well as preparation method and application thereof - Google Patents
Mussel mucoprotein liquid product as well as preparation method and application thereof Download PDFInfo
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- CN103520766A CN103520766A CN201310441954.1A CN201310441954A CN103520766A CN 103520766 A CN103520766 A CN 103520766A CN 201310441954 A CN201310441954 A CN 201310441954A CN 103520766 A CN103520766 A CN 103520766A
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Abstract
The invention discloses a mussel mucoprotein liquid product as well as a preparation method and an application thereof. One or more of a modifier, an additive and a crosslinker are added into mussel mucoprotein to form a new product which can serve as a wound repair product, a wound protection product, a medical biological bonder product, a medical coating product, an industrial coating product, a biochemical reagent product or the like. According to the mussel mucoprotein liquid product and the preparation method of the mussel mucoprotein liquid product, the modifier, the additive and the crosslinker are added into the mussel mucoprotein, so that the stability of a protein structure is enhanced, the protein molecular weight is increased, and the mussel mucoprotein liquid product is widely applied in various fields.
Description
Technical field
The present invention relates to sea-mussel mucin fluid product, relate to specifically and use sea-mussel mucin for raw material, to prepare the method for the fluid products such as wound repair product, protecting wound surface product, medical bio adhesive product, medical coating product, industrial coating product, biochemical reagents product.
Background technology
Sea-mussel mucin (Mussel adhesive protein, MAP), is also named mussel byssus albumen (Mytilus edulis foot protein, Mefp) from seashells Mytilus edulis, is a kind of Special Proteins of its secretion.Mytilus edulis is attached on longshore reef or the bottom of steamer conventionally in a cluster, has the ability in coastal waters tolerance wave stroke.In fact Mytilus edulis almost can be attached in the substrate of any material, as metal, timber, glass etc. extremely securely.The main cause that mussel has above-mentioned characteristic is can generate and store a kind of special albumen in its byssus gland, is discharged on the surface of solids of rock one class by byssus, forms the combination of water resistant, thereby oneself is fixing.
Why sea-mussel mucin has extremely strong adhesive effect, key be to be rich in its structure 3,4 dihydroxyphenylalanines (DOPA), the phenolic groups of DOPA has very strong metal-chelating ability, at material surface, form irreversible metal-organic complex, form very strong hydrogen bonded also and between protein isopolarity polymer.In addition,, in byssus adhesive curing process, the oxidation of part DOPA residue generates ortho position diquinone, by producing covalent cross-linking between additive reaction and lysine and cysteine, has further increased its cohesiveness.
At present, although sea-mussel mucin has above feature, form the considerably less of product application.Commercial sea-mussel mucin product only has the Cell-Tak of BD Biosciences company, and this product is directly used with sea-mussel mucin solution state, for field of cell culture.The cell adhesion that sea-mussel mucin product is few, application only limits to microcosmic, illustrates that sea-mussel mucin has lost some natural attribute in its acquisition process, has limited its use.
In addition, the method of disclosed several separating and purifying sea-mussel mucins in China Patent Publication No. (CN101348520, CN101348518, CN101585874) and PCT international application no (PCT/CN2012/073167), at this, quote, all belong to the content that the present invention records.
Summary of the invention
The object of the present invention is to provide preparation method, sea-mussel mucin fluid product and the application thereof of sea-mussel mucin fluid product, take sea-mussel mucin as raw material, modes such as adopting modification, add or be cross-linked forms new sea-mussel mucin fluid product, to solve in prior art, with sea-mussel mucin solution state, directly uses the defect of being brought.
For achieving the above object, the application proposes a kind of preparation method of sea-mussel mucin fluid product, comprises the following steps:
Step 1: obtain or prepare liquid sea-mussel mucin;
Step 2: add dressing agent in described liquid sea-mussel mucin, described sea-mussel mucin is carried out to chemical modification.
Preferably, the method, after described step 2, also comprises step 3: in the sea-mussel mucin after modifying, add cross-linking agent, by described cross-linking agent, make described sea-mussel mucin form intramolecular crosslinking.
Preferably, the method, after described step 2, also comprises step 3 ': in the sea-mussel mucin after modifying, add additive and cross-linking agent, by described cross-linking agent, make described sea-mussel mucin and described additive form intermolecular cross-linking.
After described step 2, also comprise: step 3 ", in the sea-mussel mucin after modifying, add additive, make to form intermolecular cross-linking between described sea-mussel mucin and described additive.
Preferably, in step 1, described liquid sea-mussel mucin is dissolved and prepares by acidity or neutral solution by sea-mussel mucin dry powder.
Preferably, described dressing agent comprises one of them or its combination of oxygen, ozone, hydrogen peroxide, iodine and preparation thereof, fluorine, chlorine, periodate, permanganate, nitrate, acetate, persulfate, fluoride.
Preferably, the adding proportion of described dressing agent is to add 0.001-50g dressing agent in every hundred milliliters of liquid sea-mussel mucins.
Preferably, the reaction pH of described dressing agent is 1.0-7.0.
Preferably, the response time of described dressing agent is 5-60min.
Preferably, described additive is one of them or combination in any of gelatin, collagen protein, hyaluronate, chitin, albumin, casein or chitosan.
Preferably, the adding proportion of described additive is to add additive 0-80g in every hundred milliliters of liquid sea-mussel mucins.
PH when preferably, described additive adds is 1.0-7.0.
Preferably, described cross-linking agent is homobifunctional agent or heterobifunctional agent.
Preferably, described homobifunctional agent is glutaraldehyde; Described heterobifunctional agent is chloropropylene oxide, Isosorbide-5-Nitrae-dihydroxy glycidyl ether, N-succinimido 3-(2-pyridine radicals two sulfur) propionic ester or succinimido-4-(N-methyl maleimide) thiacyclohexane-1-carbonic ester.
Preferably, the adding proportion of described cross-linking agent is to add cross-linking agent 0-50g in every hundred milliliters of liquid sea-mussel mucins.In addition, the application also proposes the described prepared sea-mussel mucin fluid product of preparation method.
In addition, the application has also proposed above-mentioned sea-mussel mucin fluid product in the application in culture vessel for wound repair product, protecting wound surface product, medical bio adhesive product, medical coating product, industrial coating product, biochemical reagents product, medical product, sterile products and cell culture.
The preparation method of sea-mussel mucin fluid product provided by the present invention and sea-mussel mucin fluid product, sea-mussel mucin is carried out to chemical modification, and it is cross-linked, strengthened the stability of protein structure, increased protein molecular weight, it can be applied even more extensively in each field.
Accompanying drawing explanation
Fig. 1 is the process sequence diagram of preparation method first embodiment of sea-mussel mucin fluid product of the present invention;
Fig. 2 is the process sequence diagram of preparation method second embodiment of sea-mussel mucin fluid product of the present invention;
Fig. 3 is the process sequence diagram of preparation method the 3rd embodiment of sea-mussel mucin fluid product of the present invention;
The process sequence diagram of preparation method the 4th embodiment that Fig. 3 ' is sea-mussel mucin fluid product of the present invention;
Fig. 4 adds the microscope figure that modifies sea-mussel mucin culture plate in embodiment 1;
Fig. 5 is the microscope figure that does not add sea-mussel mucin culture plate in embodiment 1;
Fig. 6 adds the microscope figure that modifies sea-mussel mucin culture plate in embodiment 2;
Fig. 7 is the microscope figure that does not add sea-mussel mucin culture plate in embodiment 2;
Fig. 8 is the implantation body's scanning electron microscope (SEM) photograph that does not add sea-mussel mucin in embodiment 3;
Fig. 9 adds implantation body's scanning electron microscope (SEM) photograph of modifying sea-mussel mucin in embodiment 3;
Figure 10 adds the microscope figure that modifies sea-mussel mucin culture plate in embodiment 6;
Figure 11 is the microscope figure that does not add sea-mussel mucin culture plate in embodiment 6;
Figure 12 is the artificial bone cell proliferation experiment contrast schematic diagram that adds and do not add sea-mussel mucin in embodiment 6.
The specific embodiment
Sea-mussel mucin can solidify and form bonding in sea water, and this feature is that existing binding agent is all incomparable, can be used to prepare the products such as corrosion-inhibiting coating; In addition, sea-mussel mucin does not have toxic action, does not cause that Human immune responses, the present invention utilize sea-mussel mucin These characteristics and sea-mussel mucin is carried out to necessary modification or interpolation yet, makes it have better attribute, forms more product.Prepared by the present invention take medical treatment and the industrial sea-mussel mucin liquid form product that sea-mussel mucin is raw material.As wound repair product, protecting wound surface product, medical bio adhesive product, medical coating product, industrial coating product, biochemical reagents product, medical product, sterile products, culture vessel etc. for cell culture.Can be used for the reparation of the various mucocutaneous wound surface such as burn, ulcer, chilblain, decubital ulcer, operation; The implantation of artificial bone, cartilage frame, periosteum, artificial joint, dentistry implant, shutoff support, spine cage, vertebral spacing device, organ sticking patch etc.; The protection of the various mucocutaneous wound surface such as burn, ulcer, chilblain, decubital ulcer, operation; The closure of operative incision; That fractures is bonding; Mucosa bonding; The body implant coatings such as artificial bone, cartilage frame, periosteum, artificial joint, dentistry implant, shutoff support, spine cage, vertebral spacing device, organ sticking patch; Cell and tissue culture; The anticorrosion of boats and ships, electronic equipment, pipeline etc.
The effect of sea-mussel mucin itself is no doubt main, and the performance of dosage form to effect, also plays positive role under certain condition.Liquid preparation is a kind of common dosage form, has following characteristics: material is dispersed in medium with molecule or graininess, and dispersion is large, absorption is fast, can play a role rapidly, and embodiment is various, bioavailability is high.
Yet, take protecting wound surface product as example, sea-mussel mucin fluid sample is sprayed directly on on wound; can form thin film at normal temperatures; there is waterproof, the characteristic such as ventilative, but directly the crosslinked film obtaining is thinner and elasticity is poor due to sea-mussel mucin, cannot play the effect of Wound protection.Liquid sea-mussel mucin is modified to the film thickness forming after processing moderate and there is certain elasticity, can meet the effect of protecting wound surface.
The present invention be take sea-mussel mucin liquid as raw material; modes such as adopt modifying, add or be crosslinked forms new sea-mussel mucin fluid product, and the product of formation can be used as wound repair product, protecting wound surface product, medical bio adhesive product, medical coating product, industrial coating product, biochemical reagents product, medical product, sterile products, cell culture by culture products etc.
Wherein, the chemical modification of protein refers to by chemical method transforms protein molecule on molecular level, in vitro by the side chain radical of protein because being undertaken covalently bound by manual method and some chemical groups, or change the backbone structure of protein molecule, thereby change the character of protein.Reagent for chemical modification is a lot, as glucosan, starch, Polyethylene Glycol etc.
In addition, with protein, protein being carried out to chemical modification is cross-linked proteins, and stability that can Enhancin matter structure increases protein molecular weight.With protein, protein is carried out to chemical modification and need use bi-functional cross-linking agent, bifunctional reagent is to have two response function groups, can with two Interaction of substituents of a protein, form intramolecular crosslinking, or with a certain Interaction of substituents of two different proteins, form the compound of intermolecular coupling, be often used to the chemical modification of protein.Bi-functional cross-linking agent is divided into homotype and special-shaped two kinds, as, glutaraldehyde be and plants homotype bi-functional cross-linking agent, its two aldehyde radicals can be respectively with two identical or different molecules on primary amino radical formation SchiffShi alkali, two molecules are coupled together.Special-shaped bifunctional reagent can be controlled the carrying out of cross-linking reaction, reduces or has avoided protein molecule self-polymerization and intersected polymerization, easily forms the protein product that is cross-linked with each other.Heterobifunctional agent can be selected from N-succinimido 3-(2-pyridine radicals two sulfur) propionic ester (SPDP), succinimido-4-(N-methyl maleimide) thiacyclohexane-1-carbonic ester (SMCC) etc.In liquid sea-mussel mucin, add dressing agent, cross-linking agent or additive, can strengthen some characteristic of sea-mussel mucin, it is applied even more extensively in each field.
Particularly, the present invention proposes a kind of preparation method of sea-mussel mucin fluid product, comprises the following steps (as Fig. 1):
Step 1: obtain or prepare liquid sea-mussel mucin;
Step 2: add dressing agent in described liquid sea-mussel mucin, described sea-mussel mucin is carried out to chemical modification.
Wherein, after described step 2, also can comprise: step 3: in the sea-mussel mucin after modifying, add cross-linking agent, by described cross-linking agent, make described sea-mussel mucin form intramolecular crosslinking (as Fig. 2).
And, after described step 2, also can comprise: step 3 ': in the sea-mussel mucin after modifying, add additive and cross-linking agent simultaneously, by described cross-linking agent, make described sea-mussel mucin and described additive form intermolecular cross-linking (as Fig. 3).
After described step 2, also comprise: step 3 ", in the sea-mussel mucin after modifying, add additive, make to form between described sea-mussel mucin and described additive intermolecular cross-linking (as Fig. 3 ').Wherein, between the groups such as the hydroxyl on protein molecule, amino, carboxyl, sulfydryl, can there is dehydrating condensation, oxidation cross-linked cross-linking reaction, be called albumen waste residue crosslinked.
Described sea-mussel mucin can be any one in 8 subclass: sea-mussel mucin 1-6(Mytilus edulis foot protein, mefp-1, mefp-2, mefp-3, mefp-4, mefp-5, mefp-6), and collagen protein preCoID, preCoIN.
According to the present invention, described liquid sea-mussel mucin is liquid sea-mussel mucin, or described liquid sea-mussel mucin is that dry powder state sea-mussel mucin is through dissolving the sea-mussel mucin solution obtaining.
Further, the concentration of described sea-mussel mucin solution is 0.01-300mg/mL.
Further, described liquid sea-mussel mucin can be dissolved and prepare by acidity or neutral solution by sea-mussel mucin dry powder.
Further, acidity or neutral solution pH value 1.0-7.0, comprising: mineral acid, example hydrochloric acid, phosphoric acid, nitric acid, boric acid, perchloric acid, periodic acid, permanganic acid etc.; Organic acid, as: formic acid, benzoic acid, acetic acid, ethanedioic acid, citric acid etc.; The buffer of weak acid and salt thereof: acetic acid-acetate buffer, phosphoric acid-phosphate buffer, boric acid-borate, citric acid-citrate etc.
According to the present invention, described dressing agent comprises: oxygen (oxygen, ozone, hydrogen peroxide), iodine and preparation thereof (iodine crystal, iodine solution, povidone iodine, iodine tincture, povidone iodine, iodoform ethereal solution etc.), fluorine, chlorine, periodate, permanganate, nitrate, acetate, persulfate, fluoride, but be not limited to this.
Further, with respect to liquid sea-mussel mucin, the adding proportion of described dressing agent is to add 0.001-50g dressing agent in every hundred milliliters of liquid sea-mussel mucins, preferably 0.01-10g.
Further, the reaction pH of described dressing agent is 1.0-7.0, preferably pH3.0-6.0.
Further, the response time of described dressing agent is arbitrarily, preferably 5-60min.
Further, the reaction temperature of described dressing agent is arbitrarily, preferably 10-50 ℃.
Further, described dressing agent can be removed, and also can not remove.
Further, described dressing agent is removed, and can adopt dialysis, also can adopt desalting column.
According to the present invention, described additive can be gelatin, collagen protein, hyaluronate, chitin, albumin, casein, and chitosan etc., but be not limited to this.
Further, with respect to liquid sea-mussel mucin, the adding proportion of described additive is to add additive 0-80g in every hundred milliliters of liquid sea-mussel mucins, preferably 1-30g.
Further, the pH during interpolation of described additive is 1.0-7.0, preferably pH3.0-6.0.
Further, the incorporation time of described additive is arbitrarily, preferably 5-60min.
Further, during the mixing of described additive, temperature is arbitrarily, preferably 10-50 ℃.
According to the present invention, the crosslinked cross-linking agent of described sea-mussel mucin and additive can be that homobifunctional agent can be also heterobifunctional agent.
Further, homobifunctional agent can be glutaraldehyde etc., but is not limited to this.
Further, heterobifunctional agent can be chloropropylene oxide, Isosorbide-5-Nitrae-dihydroxy glycidyl ether, N-succinimido 3-(2-pyridine radicals two sulfur) propionic ester (SPDP), succinimido-4-(N-methyl maleimide) thiacyclohexane-1-carbonic ester (SMCC) etc., but be not limited to this.
Further, with respect to liquid sea-mussel mucin, the adding proportion of described cross-linking agent is to add cross-linking agent 0-50g in every hundred milliliters of liquid sea-mussel mucins, preferably 1-10g.
Further, the reaction pH of described cross-linking agent is 1.0-7.0, preferably pH3.0-6.0.
Further, the response time of described cross-linking agent is arbitrarily, preferably 5-60min.
Further, the reaction temperature of described cross-linking agent is arbitrarily, preferably 10-50 ℃.
Further, described cross-linking agent agent can be removed, and also can not remove.
Further, described cross-linking agent is removed, and can adopt dialysis, also can adopt desalting column.
In addition, the present invention proposes by the prepared sea-mussel mucin fluid product of above-mentioned preparation method.
The same, the present invention has also proposed above-mentioned prepared sea-mussel mucin fluid product in the application in culture vessel for wound repair product, protecting wound surface product, medical bio adhesive product, medical coating product, industrial coating product, biochemical reagents product, medical product, sterile products and cell culture.
Specific embodiment:
Get concentration and be 1 milliliter of the sea-mussel mucin solution of 0.01mg/mL, logical oxygen (modifications) 30min under room temperature condition, the albumen after modified is used for BEK-21 cell culture, at cell, inoculates latter 6 hours observed results.As shown in Figure 4, Figure 5, do not compare with adding the result of sea-mussel mucin, add the culture plate of sea-mussel mucin after modifying, cell attachment is more, proves the adherent performance that has promoted cell adding of sea-mussel mucin.
The protein solution that identical preparation method obtains is for animal bone extirpation experiment, and be about to sea-mussel mucin and bone meal and adjust after mixing and join the damaged place of rat back bone, with the bone meal that do not add sea-mussel mucin in contrast, the six weeks damaged place of rear observation osteogenesis situations.Can see: the bone meal that does not add albumen is compared, add the bone meal of sea-mussel mucin and can obviously find out the generation that has blood vessel after growing six weeks in the damaged place of bone, prove the growth that can promote that animal bone is damaged that adds of sea-mussel mucin.
1 milliliter of sea-mussel mucin solution getting concentration and be 5mg/mL, it is 0.01%(mass percent that room temperature condition adds concentration) iodine solution 100 μ L, standby after sea-mussel mucin is fully mixed homogeneously with iodine solution.Albumen after modified, for HEK cell culture, is inoculated latter 2 hours observed results at cell.
As shown in Figure 6, Figure 7, do not compare with adding the result of sea-mussel mucin, add the culture plate of sea-mussel mucin after modifying, cell attachment is more, proves the adherent performance that has promoted cell adding of sea-mussel mucin.
1 milliliter of sea-mussel mucin solution getting concentration and be 5mg/mL, it is 10%(mass percent that room temperature condition adds concentration) iodine solution 100 μ L, after sea-mussel mucin is fully mixed homogeneously with iodine solution, room temperature removes after unnecessary iodine standby.Albumen after modified is for BHK-21 cell culture in implantation body, after cell is inoculated latter 24 hours, scanning electron microscopic observation result as shown in Figure 8, Figure 9: do not compare with adding the result of sea-mussel mucin, add in the implantation body of sea-mussel mucin after modifying, Growth of Cells is more, proves the adherent growth performance that has promoted cell adding of sea-mussel mucin.
1 milliliter of sea-mussel mucin solution getting concentration and be 300mg/mL, it is 5%(mass percent that room temperature condition adds concentration) iodine solution 100 μ L, sea-mussel mucin is fully mixed homogeneously with iodine solution, room temperature removes after excess iodine standby.Albumen after modified is bonding for rabbit skin incision, and result is as follows:
Chemical adhesive, when bonding rabbit skin incision, is easily burnt and otch surrounding skin is unfavorable for healing, and the bonding otch surrounding skin softness of mussel adh esive protein, healing rate is fast.
Get concentration and be 1 milliliter of the sea-mussel mucin solution of 250mg/mL, it is 5%(mass percent that room temperature condition adds concentration) iodine solution 100 μ L, sea-mussel mucin is fully mixed homogeneously with iodine solution, room temperature adds 10%(mass percent after removing excess iodine wherein) gelatin, after mix homogeneously, obtain new binder solution.The sea-mussel mucin solution of getting new binder solution and not adding gelatin (only modify) carries out determining bonding strength experiment, is about to two kinds of protein solutions and is coated onto respectively on dry Corii Caprae seu Ovis, measures the lap shear strength of two kinds of albumen.
The lap shear strength of two kinds of protein solutions
| Solution | Lap shear strength (MPa) |
| Sea-mussel mucin after modifying | 0.01 |
| After modifying, sea-mussel mucin adds gelatin | 0.015 |
Can find out, after interpolation gelatin, the adhesive strength of albumen can strengthen.
1 milliliter of sea-mussel mucin solution getting concentration and be 5mg/mL, it is 1%(mass percent that room temperature condition adds concentration) copperas solution 10 μ L, standby after sea-mussel mucin is fully mixed homogeneously with copperas solution.Albumen after modified, for MSC cell culture, is inoculated latter 2 hours observed results at cell as follows:
As shown in Figure 10, Figure 11, do not compare with adding the result of sea-mussel mucin, add the culture plate of sea-mussel mucin after modifying, cell attachment is more, proves the adherent performance that has promoted cell adding of sea-mussel mucin.
Albumen after modification for artificial bone cell proliferation experiment, contrasts with the cultivation effect of domestic bone meal, import bone meal respectively, result as shown in figure 12, wherein, series 1: domestic bone meal; Series 2: domestic bone meal adds modifies sea-mussel mucin; Series 3: import bone meal; Series 4: domestic bone meal adds unmodified sea-mussel mucin.
As seen from Figure 12, at different growth natural law, the cell proliferation situation on the bone meal of sea-mussel mucin of adding is all better than not adding domestic bone meal and the import bone meal of albumen, proves that sea-mussel mucin can promote the propagation of osteocyte.
Get concentration and be 1 milliliter of the sea-mussel mucin solution of 5mg/mL; it is 1%(mass percent that room temperature condition adds concentration) copperas solution 10 μ L; after sea-mussel mucin is fully mixed homogeneously with copperas solution; add under 5% glutaraldehyde solution room temperature crosslinked 2 hours; protein solution after crosslinked is at room temperature dry can form transparent, waterproof, breathable films, can be used as protecting wound surface material for the protection of wound surface.
Get concentration and be 1 milliliter of the sea-mussel mucin solution of 5mg/mL, it is 1%(mass percent that room temperature condition adds concentration) copperas solution 10 μ L, after sea-mussel mucin is fully mixed homogeneously with copperas solution, add under 5% glutaraldehyde solution room temperature crosslinked 2 hours, in protein solution after crosslinked, add again 10% casein, fully stir and make mix homogeneously, the liquid forming at room temperature dry can form transparent, waterproof, breathable films, and film strength will be when additive-free, the product forming can be used as protecting wound surface material for the protection of wound surface.
The preparation method of sea-mussel mucin fluid product provided by the present invention and sea-mussel mucin fluid product, by add dressing agent, additive, cross-linking agent in sea-mussel mucin, strengthened the stability of protein structure, increased protein molecular weight, it can be applied even more extensively in each field.
Certainly; the present invention also can have other various embodiments; in the situation that not deviating from spirit of the present invention and essence thereof; those of ordinary skill in the art can make according to the present invention various corresponding changes and distortion, but these corresponding changes and distortion all should belong to the protection domain of the claims in the present invention.
Claims (17)
1. a preparation method for sea-mussel mucin fluid product, is characterized in that, comprises the following steps:
Step 1: liquid sea-mussel mucin is provided;
Step 2: add dressing agent in described liquid sea-mussel mucin, described sea-mussel mucin is carried out to chemical modification.
2. the preparation method of sea-mussel mucin fluid product according to claim 1, is characterized in that, after described step 2, also comprises:
Step 3: add cross-linking agent in the sea-mussel mucin after modifying, make described sea-mussel mucin form intramolecular crosslinking by described cross-linking agent.
3. the preparation method of sea-mussel mucin fluid product according to claim 1, is characterized in that, after described step 2, also comprises:
Step 3 ", in the sea-mussel mucin after modifying, add additive, make to form intermolecular cross-linking between described sea-mussel mucin and described additive.
4. the preparation method of sea-mussel mucin fluid product according to claim 1, is characterized in that, after described step 2, also comprises:
Step 3 ': in the sea-mussel mucin after modifying, add additive and cross-linking agent, by described cross-linking agent, make described sea-mussel mucin and described additive form intermolecular cross-linking.
5. according to the preparation method of the sea-mussel mucin fluid product described in claim 1 or 2 or 3 or 4, it is characterized in that, in step 1, described liquid sea-mussel mucin is liquid sea-mussel mucin, or described liquid sea-mussel mucin is that dry powder state sea-mussel mucin is through dissolving the sea-mussel mucin solution obtaining.
6. according to the preparation method of the sea-mussel mucin fluid product described in claim 1 or 2 or 3 or 4, it is characterized in that, described dressing agent comprises one of them or its combination of oxygen, ozone, hydrogen peroxide, iodine and preparation thereof, fluorine, chlorine, periodate, permanganate, nitrate, acetate, persulfate, fluoride.
7. according to the preparation method of the sea-mussel mucin fluid product described in claim 1 or 2 or 3 or 4, it is characterized in that, the adding proportion of described dressing agent is to add 0.001-50g dressing agent in every hundred milliliters of liquid sea-mussel mucins.
8. according to the preparation method of the sea-mussel mucin fluid product described in claim 1 or 2 or 3 or 4, it is characterized in that, the reaction pH of described dressing agent is 1.0-7.0.
9. according to the preparation method of the sea-mussel mucin fluid product described in claim 1 or 2 or 3 or 4, it is characterized in that, the response time of described dressing agent is 5-60min.
10. according to the preparation method of the sea-mussel mucin fluid product described in claim 3 or 4, it is characterized in that, described additive is one of them or combination in any of gelatin, collagen protein, hyaluronate, chitin, albumin, casein or chitosan.
11. according to the preparation method of the sea-mussel mucin fluid product described in claim 3 or 4, it is characterized in that, the adding proportion of described additive is to add additive 0-80g in every hundred milliliters of liquid sea-mussel mucins.
12. according to the preparation method of the sea-mussel mucin fluid product described in claim 3 or 4, it is characterized in that, pH when described additive adds is 1.0-7.0.
13. according to the preparation method of the sea-mussel mucin fluid product described in claim 2 or 4, it is characterized in that, described cross-linking agent is homobifunctional agent or heterobifunctional agent.
The preparation method of 14. sea-mussel mucin fluid products according to claim 11, is characterized in that, described homobifunctional agent is glutaraldehyde; Described heterobifunctional agent is chloropropylene oxide, Isosorbide-5-Nitrae-dihydroxy glycidyl ether, N-succinimido 3-(2-pyridine radicals two sulfur) propionic ester or succinimido-4-(N-methyl maleimide) thiacyclohexane-1-carbonic ester.
15. according to the preparation method of the sea-mussel mucin fluid product described in claim 2 or 4, it is characterized in that, the adding proportion of described cross-linking agent is to add cross-linking agent 0-50g in every hundred milliliters of liquid sea-mussel mucins.
16. by the prepared sea-mussel mucin fluid product of preparation method described in claim 1 or 2 or 3 or 4.
The sea-mussel mucin fluid product of 17. claim 16 is in the application in culture vessel for wound repair product, protecting wound surface product, medical bio adhesive product, medical coating product, industrial coating product, biochemical reagents product, medical product, sterile products and cell culture.
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Cited By (27)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104323927A (en) * | 2014-11-06 | 2015-02-04 | 济南星河康泰贸易有限公司 | Application of mussel adhesive protein in preparing cosmetics for skin beautifying |
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